Fig. 1

aPali is anchored to the membrane and inhibits RSV in cells. a Schematic of aPali anchored to the plasma membrane. b Schematic of aPali and sPali mRNA delivery and expression. c Cells were transfected with vehicle control or 1 μg of either aPali heavy chain only, aPali light chain, or both the heavy and light chain mRNAs. 24 h later, cells were fixed, permeabilized, and stained with a donkey anti-human secondary antibody (white). Scale bar represents 10 μm. d Cells were transfected with 1 μg of either aPali or sPali mRNA. At 2, 4, 6, 12, and 24 h, cells were fixed, permeabilized or not, and stained for the expressed antibody (white). Scale bar represents 10 μm. e Cells were transfected with vehicle control or 1 μg of either aPali heavy chain only, aPali light chain only, or both the heavy and light chain mRNAs. After overnight incubation, cells were infected for 24 h before being fixed and stained for RSV N (green), panRSV (blue), and aPali (red). Scale bar represents 10 μm. f Quantification of the mean volume of the RSV N signal per cell from microscopy images in part (e). Error bars represent 95% confidence intervals. Asterisks indicate p < 0.0001 (Mann–Whitney U test). 25 cells were analyzed per group. Results represent mean of two independent experiments. g Cells were transfected with vehicle control or 1 μg of aPali and infected with RSV at MOI of 0.01, 0.1, and 1. Supernatants were collected at 24 hpi and virion titers were measured by plaque assay. Each data point represents mean of 4 replicate wells. Error bars represent standard deviation. Dotted line represents lower limit of detection. Asterisks indicate p < 0.0001 (one-way ANOVA of log-transformed data). Results represent mean of three independent experiments